Protecting plants against the fungal pathogen Sclerotinia sclerotiorum using host-induced gene silencing
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Abstract
Sclerotinia sclerotiorum is a fungal pathogen that affects Canadian crops every year. Brassica napus (canola) is particularly vulnerable. Chemical fungicides are widely used to control S. sclerotiorum. However, with increasing concerns about resistance and ecological effects, it is necessary to develop new and improved control methods. Host-induced gene silencing (HIGS) is an RNA interference technology that has the potential to be harnessed to create transgenic, pathogen-resistant crops. In this study, I analyzed the effectiveness of four independently transformed lines of Arabidopsis thaliana expressing RNA to silence the NOB1 gene (SS1G_07873) in S. sclerotiorum. Three of the lines expressed double-stranded (ds)RNA while the fourth line expressed paperclip (pc)RNA, a novel RNA structure which folds over itself on both ends to protect it from nucleases. The analysis was done through detached-leaf lesion assays and transcript knockdown analysis of the NOB1 target gene as well as three known interacting genes. Though improvements are required, the lesion assays show significant decreases in lesion size. Transcript knockdown analysis revealed insignificant changes in NOB1 expression. The interacting genes analyzed showed a significant reduction in transcript abundance, indicating that further experiments are required to understand the processing of foreign RNA in the plant. With further testing to optimize HIGS using SS1G_07873, this technology has the potential to be replicated in crop plants as a means of protection against S. sclerotiorum.